Electrical Stimulation of C6 Glia-Precursor Cells In Vitro Differentially Modulates Gene Expression Related to Chronic Pain Pathways

Abstract

Glial cells comprise the majority of cells in the central nervous system and exhibit diverse functions including the development of persistent neuropathic pain. While earlier theories have proposed that the applied electric field specifically affects neurons, it has been demonstrated that electrical stimulation (ES) of neural tissue modulates gene expression of the glial cells. This study examines the effect of ES on the expression of eight genes related to oxidative stress and neuroprotection in cultured rodent glioma cells. Concentric bipolar electrodes under seven different ES types were used to stimulate cells for 30 min in the presence and absence of extracellular glutamate. ES consisted of rectangular pulses at 50 Hz in varying proportions of anodic and cathodic phases. Real-time reverse-transcribed quantitative polymerase chain reaction was used to determine gene expression using the ∆∆Cq method. The results demonstrate that glutamate has a significant effect on gene expression in both stimulated and non-stimulated groups. Furthermore, stimulation parameters have differential effects on gene expression, both in the presence and absence of glutamate. ES has an effect on glial cell gene expression that is dependent on waveform composition. Optimization of ES therapy for chronic pain applications can be enhanced by this understanding.

Modulation of microglial activation states by spinal cord stimulation in an animal model of neuropathic pain: Comparing high rate, low rate, and differential target multiplexed programming

“…14,15 It is known that spinal cord stimulation (SCS) can modulate transcriptomic expression of neural cells in animal models. 8,9,12,13,[32][33][34][35][36] More specifically, our work demonstrated that changes in SCS stimulation parameters could differentially modulate gene expression of neurons and glial cells by using an approach with multiplexed electrical signals (called differential target multiplexed programming, DTMP). DTMP can modulate biological processes associated with neuron-glial interactions more effectively than other standard SCS approaches using high rate or low rate programs (HRP or LRP), while providing significant relief from thermal and mechanical hypersensitivity, which was also significantly improved relative to these standard SCS treatments.…”

“… 10 , 11 Recently, our lab and others have shown, using transcriptomics and proteomics in animal models, that neuropathic pain induces a complex response composed of metabolic, inflammatory, and immune biological processes involved in neuron-glial interactions that SCS can modulate. 8 , 9 , 12 , 13 …”

Neurovascular-modulation

“…Activation of neurogenic regulation of cardiac function [109][110][111] or brain clearance [112] including electrical stimulation of perivascular innervation [113] is distinct from the direct BBB polarization of Principle 2. Electrical stimulation of glia [114][115][116] and subsequent astrocyte regulation of the BBB [117] are also parallel but distinct pathways.…”

Synthesis of Polycyclic Ether-Benzopyrans and In Vitro Inhibitory Activity against Leishmania tarentolae

“…Brain heart infusion media (BHI) supplemented with penicillin, streptomycin, and hemin was used for L. tarentolae culturing following the method of Morgenthaler et al [ 97 ]. Axenic Rattus norvegicus C6 glioma cells (ATCC CCL-107) were cultured in FALCON 6-well culture plates using high glucose Dulbecco’s Modified Eagles Medium (incomplete DMEM; Sigma Life Sciences D6429; St. Louis, MO, USA) supplemented with 15% ( v / v ) horse serum (ATCC; Manassas, VA, USA) and 5% ( v / v ) heat-treated fetal bovine serum (GIBCO; Waltham, MA, USA) (now designated as complete DMEM) at 37 °C and maintained under a 5% CO 2 atmosphere following the method of Vallejo et al [ 98 ]. For experimentation, cells were harvested by trypsinization, following the manufacturer’s recommendations (Sigma Aldrich), and resuspended in complete DMEM.…”